胰腺癌是一種致死率極高的惡性疾病,在全世界所有的癌症中死亡率排列第八, 在美國排列第四。 很多因素造成了胰腺癌較差的預後,其中包括: 早期檢出率極低; 較少胰腺癌患者的腫瘤適宜手術切除;高轉移率;以及對傳統放療和化療具有較高抗性等。 因此,發展新的治療藥物迫在眉睫。 / 近年來, 植物藥以及從這些植物藥裡分離出的天然化合物, 單獨使用或者與傳統化療藥物合併使用時, 都顯示出對不同類型的癌症具有較好療效。植物藥毛萼香茶菜(唇形科)含有豐富的具有抗癌活性的二萜類化合物。其中毛萼乙素(EriB) 是一個擁有最好抗癌活性的對映-貝殼杉烷型二萜化合物。 基於此背景, 本研究的目標為:利用胰腺癌體外體內模型, 研究EriB的抗胰腺癌活性以及誘導胰腺癌細胞凋亡的機理。 / 體外實驗中, EriB對四種胰腺癌細胞株都顯示了顯著的細胞毒活性,其活性與化療藥物喜樹堿類似。其中, EriB對胰腺癌細胞株CAPAN-2活性最強, 半數致死濃度IC₅₀為0.73 μM。細胞凋亡特徵:細胞核凝聚, 磷脂醯絲氨酸外翻, DNA梯狀條帶以及片斷化,在EriB誘導的胰腺癌細胞株CAPAN-2中出現。此外, EriB還造成癌細胞在細胞週期G2/M期的阻滯。機理研究發現, EriB是通過啟動絲裂原活化蛋白激酶(MAPK), caspase及 p53信號通路來誘導細胞凋亡和細胞週期阻滯的。抗凋亡蛋白與促凋亡蛋白比率(bcl-2/bak)的減少也可能對啟動細胞凋亡內途徑發揮一定作用。除此以外, EriB對癌細胞的細胞毒活性及致凋亡作用依賴于活性氧分子(ROS)的產生。在對細胞進行抗氧化劑預處理的實驗中發現, 只有含巰基基團的抗氧化劑能夠有效的阻斷EriB對癌細胞的活性。進一步實驗證明, EriB對細胞內兩個抗氧化系統: 谷胱甘肽系統及硫氧還蛋白系統的抑制作用導致了ROS在癌細胞中的積聚。同時,ROS的產生啟動了MAPK,熱休克蛋白70以及caspase信號通路,卻抑制了NFκB通路。 / 動物體內實驗證實, 每天對胰腺癌細胞移植瘤裸鼠進行腹腔注射EriB(2.5 毫克/千克),能有效的抑制腫瘤生長, 並且對心臟,肝臟和腎臟沒有引起顯著毒性。 對腫瘤組織的分析表明, 給藥組(EriB)比溶劑對照組出現更多的細胞凋亡, 並產生較多的ROS積聚。 / 綜上所述, 本項研究首次闡述了EriB具有顯著的體內外抗胰腺癌活性。機理研究證明, EriB抑制胰腺癌細胞內兩個含巰基基團的抗氧化系統, 從而導致ROS在細胞中積聚, 並啟動(或抑制)了包括MAPK, p53, caspase和NFB在內的信號通路, 最終導致癌細胞死亡。 此外, 動物體內研究證明EriB的抗腫瘤生長活性和低毒性, 令該化合物具有潛力進一步研究發展成為抗胰腺癌的新藥物。 / Pancreatic cancer is the fourth and eighth leading cause of cancer-related deaths in the U.S. and worldwide, respectively. Its poor prognosis is attributed to its late diagnosis, limitation to surgical resection, aggressive local invasion, and early metastases, as well as high resistance to chemotherapy and radiotherapy. Therefore, a search for an alternative to therapeutic agents is in desperate need. / In recent years, herbal medicines or natural compounds isolated from herbs either used alone or in combination with conventional anti-cancer agents have been shown to have beneficial effects on various cancers. In this context, the Chinese herb Isodon eriocalyx (Dunn.) Hara (family Lamiaceae) is a well-known source of anti-cancer diterpenoids, the most potent one being Eriocalyxin B (EriB, an ent-kauranoid). Therefore, the aims of the present study are to investigate the anti-tumor activities of EriB in human pancreatic adenocarcinoma cells and tumor-bearing mouse model, as well as the underlying mechanisms. / Our results showed that EriB exhibited significant cytotoxic effects on four pancreatic adenocarcinoma cell lines, with potencies being comparable to that of chemotherapeutic agent camptothecin. EriB had the most potent cytotoxicity in CAPAN-2 cells with IC₅₀ = 0.73 μM. The hallmark features of apoptosis, such as nuclear condensation, translocation of phosphatidylserine, DNA laddering, and DNA fragmentation were observed in EriB-treated CAPAN-2 cells. On the other hand, EriB also induced G2/M phase cell cycle arrest. Mechanistic studies revealed that EriB induced apoptosis and cell cycle arrest through the activation of MAPKs (p38, ERK1/2), caspase cascade, and p53/p21/cdk1-cyclinB1 signaling pathways. A decrease in the ratio of anti-apoptotic to pro-apoptotic proteins (bcl-2/bak) also contributed to the activation of intrinsic apoptotic pathway. Further investigation showed that EriB-induced cytotoxic and apoptotic effects were dependent on reactive oxygen species (ROS) production. Such demonstrated effects could be inhibited by pre-treatment with thiol-containing antioxidants. Furthermore, EriB induced ROS was mediated via the inhibition of two main antioxidant systems, namely glutathione and thioredoxin systems. EriB-mediated ROS activated multiple targets or signal pathways, including MAPK, heat shock protein (Hsp) 70, and caspase cascade, while inhibiting the NFκB pathway. / On the other hand, in vivo study demonstrated that daily intraperitoneal administration of EriB (2.5mg/kg/day) in human pancreatic tumor xenografts BALB/c nude mice significantly inhibited tumor growth, but without having toxicity in the heart, liver and kidney. In addition, EriB treatments induced in vivo cell apoptosis and superoxide production as observed in tumor tissues. / In conclusion, the present study reports for the first time that EriB has possessed anti-proliferative activities in pancreatic cancer cells. The anti-proliferative effects of EriB on CAPAN-2 cells could be attributable to the regulation of cellular apoptosis and cell cycle arrest. The inhibitory effects of EriB on two antioxidant systems result in the accumulation of ROS, which in turn activate MAPK, p53, Hsp70 and caspase cascade, while inhibiting NFB pathway and finally leading to pancreatic cancer cell death. Meanwhile, in vivo study further confirms the anti-tumor properties of EriB, suggesting that EriB could be considered as a potential chemotherapeutic agent for patients with pancreatic cancer. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Li, Lin. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 207-230). / Abstracts also in Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iv / Publications --- p.vi / Acknowledgements --- p.vii / Table of contents --- p.ix / List of figures --- p.xv / List of tables --- p.xix / List of abbreviations --- p.xx / Chapter Chapter1 --- General Introduction --- p.1 / Chapter 1.1 --- The pancreas --- p.2 / Chapter 1.1.1 --- Anatomy of the pancreas --- p.2 / Chapter 1.1.2 --- Histology of the pancreas --- p.4 / Chapter 1.1.3 --- Exocrine pancreas --- p.5 / Chapter 1.1.3.1 --- Structure of secretory acini, ducts and stroma in pancreas --- p.5 / Chapter 1.1.3.2 --- Functions of exocrine pancreas --- p.6 / Chapter 1.1.4 --- Endocrine pancreas --- p.9 / Chapter 1.1.4.1 --- Structure of islets cells --- p.10 / Chapter 1.1.4.2 --- Functions of endocrine pancreas --- p.10 / Chapter 1.1.5 --- Disorders of the pancreas --- p.11 / Chapter 1.2 --- Pancreatic cancer --- p.14 / Chapter 1.2.1 --- Epidemiology --- p.14 / Chapter 1.2.2 --- The risks and causes of pancreatic cancer --- p.15 / Chapter 1.2.3 --- Signs and symptoms of pancreatic cancer --- p.18 / Chapter 1.2.4 --- Types of pancreatic cancer --- p.19 / Chapter 1.2.5 --- Diagnosis of pancreatic cancer --- p.21 / Chapter 1.2.6 --- Staging of pancreatic cancer --- p.27 / Chapter 1.3 --- Treatments for pancreatic cancer --- p.29 / Chapter 1.3.1 --- Surgery --- p.29 / Chapter 1.3.2 --- Chemotherapy --- p.30 / Chapter 1.3.2.1 --- 5-fluorouracil (5-FU) --- p.32 / Chapter 1.3.2.2 --- Gemcitabine (Gem) --- p.33 / Chapter 1.3.2.3 --- Other cytotoxic agents --- p.34 / Chapter 1.3.3 --- Radiotherapy --- p.35 / Chapter 1.3.4 --- Target therapies --- p.37 / Chapter 1.3.4.1 --- Antiangiogenic therapy --- p.37 / Chapter 1.3.4.2 --- Epidermal growth factor receptor (EGFR) signaling inhibitors --- p.39 / Chapter 1.3.4.3 --- Hedgehog and Notch signaling pathways inhibitors --- p.41 / Chapter 1.3.5 --- Gene therapy --- p.42 / Chapter 1.3.6 --- Immunotherapy --- p.45 / Chapter 1.3.7 --- Combination therapies --- p.46 / Chapter 1.4 --- Molecular targets for pancreatic cancer chemotherapy --- p.49 / Chapter 1.4.1 --- Therapies-induced apoptosis --- p.49 / Chapter 1.4.1.1 --- Caspase cascade and bcl-2 Family --- p.49 / Chapter 1.4.1.2 --- Role of mitogen-activated protein kinases (MAPKs) in apoptosis --- p.50 / Chapter 1.4.2 --- Nuclear factor-κB activation in pancreatic cancer --- p.50 / Chapter 1.4.3 --- The PI3K and AKT pathway --- p.51 / Chapter 1.4.4 --- JAK/STAT pathway --- p.51 / Chapter 1.4.5 --- Other molecular targets --- p.52 / Chapter 1.5 --- Herbal medicine as an alternative treatment for cancer treatment --- p.53 / Chapter 1.5.1 --- Herbal medicines for different types of cancer treatment --- p.53 / Chapter 1.5.2 --- Herbal medicines for pancreatic cancer treatment --- p.59 / Chapter 1.6 --- Introduction of Isodon eriocalyx (Dunn.) Hara --- p.61 / Chapter 1.6.1 --- Background of Isodon genus and Isodon eriocalyx (Dunn.) Hara --- p.61 / Chapter 1.6.2 --- Diterpenoids from Isodon species and their activities --- p.62 / Chapter 1.6.3 --- The potential anti-cancer activity of Eriocalyxin B, a diterpenoid isolated from Isodon eriocalyx (Dunn.) Hara --- p.62 / Chapter 1.7 --- Aims and objectives of this study --- p.66 / Chapter Chapter 2 --- Eriocalyxin B induces apoptosis and cell cycle arrest in pancreatic adenocarcinoma cells through caspase- and p53-dependent pathways --- p.67 / Chapter 2.1 --- Introduction --- p.68 / Chapter 2.2 --- Materials and methods --- p.71 / Chapter 2.2.1 --- Preparation and quality control of Eriocalyxin B --- p.71 / Chapter 2.2.2 --- Materials --- p.72 / Chapter 2.2.3 --- Cell culture --- p.72 / Chapter 2.2.4 --- Preparation of human peripheral blood mononuclear cells (PBMC) --- p.73 / Chapter 2.2.5 --- Cytotoxicity assay --- p.75 / Chapter 2.2.6 --- Hoechst 33258 staining for morphological evaluation --- p.76 / Chapter 2.2.7 --- DNA fragmentation detection by DNA ladder --- p.76 / Chapter 2.2.8 --- Cell death detection ELISA --- p.77 / Chapter 2.2.9 --- Apoptosis detection by flow cytometry --- p.78 / Chapter 2.2.10 --- Cell cycle analysis by flow cytometry --- p.78 / Chapter 2.2.11 --- Western blot analysis --- p.79 / Chapter 2.2.12 --- Statistical analysis --- p.80 / Chapter 2.3 --- Results --- p.81 / Chapter 2.3.1 --- EriB induces cytotoxic effect in human pancreatic cancer cells --- p.81 / Chapter 2.3.2 --- EriB induces apoptosis in CAPAN-2 cells --- p.85 / Chapter 2.3.3 --- Activation of pro-apoptotic caspases in EriB-treated CAPAN-2 cells --- p.89 / Chapter 2.3.4 --- Modulation of bcl-2/bak ratio in EriB-treated CAPAN-2 cells --- p.92 / Chapter 2.3.5 --- EriB causes G2/M cell cycle arrest --- p.94 / Chapter 2.3.6 --- EriB modulates expression of G2/M cell cycle regulatory proteins through activation of the p53 pathway --- p.96 / Chapter 2.4 --- Discussion --- p.99 / Chapter Chapter 3 --- Eriocalyxin B induces apoptosis in pancreatic cancer CAPAN-2 cells via mediation of reactive oxygen species --- p.107 / Chapter 3.1 --- Introduction --- p.108 / Chapter 3.2 --- Materials and methods --- p.113 / Chapter 3.2.1 --- Materials --- p.113 / Chapter 3.2.2 --- Cell culture and MTT assay --- p.113 / Chapter 3.2.3 --- Apoptosis detection by flow cytometry --- p.114 / Chapter 3.2.4 --- Reactive oxygen species (ROS) detection by flow cytometry --- p.114 / Chapter 3.2.5 --- Glutathione assessment --- p.115 / Chapter 3.2.6 --- Glutathione peroxidase (GPx) activity detection --- p.116 / Chapter 3.2.7 --- Thioredoxin reductase (TrxR) activity detection --- p.116 / Chapter 3.2.8 --- Nuclear and cytosolic fractionation --- p.117 / Chapter 3.2.9 --- Western blot analysis --- p.117 / Chapter 3.2.10 --- Electrophoretic mobility shift assay --- p.119 / Chapter 3.2.11 --- Statistical analysis --- p.119 / Chapter 3.3 --- Results --- p.120 / Chapter 3.3.1 --- Thiol-containing antioxidants inhibits EriB-induced cytotoxic effects --- p.120 / Chapter 3.3.2 --- Thiol-containing antioxidants inhibits EriB-induced apoptotic effects --- p.122 / Chapter 3.3.3 --- Effects of EriB on hydrogen peroxide production --- p.125 / Chapter 3.3.4 --- EriB depletes glutathione level and suppresses GPx activity --- p.128 / Chapter 3.3.5 --- EriB inhibits thioredoxin system and activates ASK1 --- p.130 / Chapter 3.3.6 --- EriB increases Hsp70 and cleaved-PARP expression through ROS --- p.134 / Chapter 3.3.7 --- EriB inhibits NFkB pathway in CAPAN-2 cells --- p.137 / Chapter 3.4 --- Discussion --- p.142 / Chapter Chapter 4 --- In vivo study of the anti-tumor efficacy of Eriocalyxin B in human pancreatic tumor xenograft model --- p.149 / Chapter 4.1 --- Introduction --- p.150 / Chapter 4.2 --- Materials and methods --- p.154 / Chapter 4.2.1 --- Establishment of a subcutaneous pancreatic cancer xenograft model --- p.154 / Chapter 4.2.2 --- Evaluation of the effects of EriB on tumor growth --- p.155 / Chapter 4.2.2.1 --- Pilot study for EriB and camptothecin treatment --- p.155 / Chapter 4.2.2.2 --- Confirmation study of effective dose of EriB --- p.156 / Chapter 4.2.2.3 --- Dose-comparison study of CPT-11 --- p.156 / Chapter 4.2.2.4 --- Comparison study of EriB and CPT-11 treatments --- p..157 / Chapter 4.2.3 --- Measurement of plasma-specific enzyme levels --- p.157 / Chapter 4.2.4 --- Assays of terminal deoxytransferase-catalyzed DNA nick-end labeling (TUNEL) --- p..158 / Chapter 4.2.5 --- Histological evaluation --- p.159 / Chapter 4.2.6 --- Detection of superoxide by DHE staining --- p.159 / Chapter 4.2.7 --- Establishment of an orthotopic model (SW1990) of pancreatic cancer and detection of the plasma biomarker CA19-9 --- p.160 / Chapter 4.2.7.1 --- Detection of CA19-9 expression by immunofluorescent staining and western blot --- p.161 / Chapter 4.2.7.2 --- Establishment of an orthotopic pancreatic cancer xenograft model by SW1990 cells --- p.162 / Chapter 4.2.8 --- Statistical analysis --- p.164 / Chapter 4.3 --- Results --- p.165 / Chapter 4.3.1 --- EriB inhibits the growth of CAPAN-2 human pancreatic tumor xenografts --- p.165 / Chapter 4.3.2 --- EriB treatments induce cell apoptosis in tumor tissues --- p.173 / Chapter 4.3.3 --- Toxicity tests for EriB --- p.175 / Chapter 4.3.3.1 --- Plasma enzyme levels after EriB treatments --- p.175 / Chapter 4.3.3.2 --- No apparent alterations in histology of the heart, liver and kidney tissues --- p..176 / Chapter 4.3.4tEriB induces superoxide production in the tumor tissues --- p.178 / Chapter 4.3.5 --- Successful establishment of an orthotopic xenograft model --- p.180 / Chapter 4.4 --- Discussion --- p.184 / Chapter Chapter 5 --- General Discussion --- p.188 / Chapter 5.1 --- Discussion --- p.189 / Chapter 5.2 --- Conclusion --- p.204 / Chapter 5.3 --- Limitations of the study --- p.205 / Chapter 5.4 --- Future work --- p.206 / Chapter Chapter 6 --- References --- p.207

Made available in DSpace on 2014-07-29T16:48:33Z (GMT). No. of bitstreams: 1 Tese Julierme_IQ_UFG_2012.pdf: 1572042 bytes, checksum: c5e12770e49c6a65ee23600c84379152 (MD5) Previous issue date: 2012-04-13 / Cerrado is the second largest Brazilian biome, occupying about 24% of the country. However, there are only 61% of preserved Cerrado areas. Therefore, studies are needed in order to preserve these natural resources. For this purpose the reassessment of systematic relationships between 88 taxa of Hyptis Jacq. were based on the vegetative and reproductive morphological characters (chapter 1). In order to perform a phenetic study, multivariate statistical analysis were used for determination of taxa distribution patterns. The relationships between the examined taxa have been expressed as Ward hierarchical clustering using multiple correspondence analysis of 50 characters distributed into 168 character states. The perceptual map shows a subdivision of Hyptis into two major clusters. Cluster I contained 41 taxa belonging to sections Cyanocephalus, Cyrta, Gymneia, Mesosphaeria, Polydesmia and Tricosphaeria which mainly showed elongated, elliptic, linear bracts, curvature of the calyx's tube upper limb, oblique calyx tube orifice, as well as smooth nutlets. Cluster II contained 47 taxa from sections Apodotes, Eriosphaeria, Hyptis, Induratae, Pachyphyllae, Pusilae and Xylodontes. Their main morphological characteristics included hemispheric or subglobose cymes, the presence of a stylopodium, slightly rough nutlets and nutlets truncate at apex. Cluster analysis focused on 6 of the morphological characters which were distinguished by discriminant correspondence analysis, revealed similar traits. Multivariate analyses were also applied in the essential oil chemovariations of 13 Hypenia species (chapter 2). The results revealed the presence of two taxonomic clusters. Cluster I included 3 species belonging to section Densiflorae in addition to H. subrosea Harley and H. aristulata (Epl.) R. Harley, and showed the highest percentages of α-muurolol (5.85 ± 3.08%). In Cluster II, which contained 8 species belonging to section Laxiflorae, the major discriminant constituents were (E)-caryophyllene (7.09 ± 4.88%), germacrene D (18.1 ± 11.4%) and bicyclogermacrene (6.65 ± 1.19%). All essential oils showed a predominance of sesquiterpenes, such as spathulenol (4.5-31.6%), caryophyllene oxide (2.2-14.4%) and selin-11-en-4α-ol (0-34.8%). Furthermore, identical clusters were revealed by multivariate analysis of chemical constituents based on carbon skeletons, as well as on 18 morphological leaf characters of the species studied. / O Cerrado é a segunda maior formação vegetal brasileira, ocupando 24% do território nacional. No entanto, restam apenas 61% de áreas preservadas de Cerrado. Desta forma, estudos para caracterização e preservação desta riqueza natural são necessários. Com este intuito, reavaliou-se a relação sistemática entre 88 espécies de Hyptis Jacq. com base em caracteres morfológicos e reprodutivos (capítulo 1). A análise estatística multivariada foi aplicada para determinar o padrão de distribuição dos taxa com o objetivo de se realizar uma análise fenética. As relações entre os taxa estudados foram expressas pelo agrupamento hierárquico de Ward utilizando a análise de correspondência múltipla com base em 50 caracteres distribuídos em 168 estados dos caracters. O mapa perceptual mostrou a subdivisão de Hyptis em duas grandes classes. A classe I foi formada por 41 taxa pertencentes às seções Cyanocephalus, Cyrta, Gymneia, Mesosphaeria, Polydesmia e Tricosphaeria, as quais demostraram brácteas linares alongadas e elípticas, curvatura do tubo do cálice acima do limbo, orifício do tubo do cálice oblíquo, bem como núculas suaves. A classe II contou com 47 taxa das seções Apodotes, Eriosphaeria, Hyptis, Induratae, Pachyphyllae, Pusilae e Xylodontes. Suas principais características morfológicas incluíram cimas hemisféricas ou subglobosas, presença de um estilopódio, núculas levemente rugosas e núculas truncadas no ápice. A análise de agrupamento focada em 6 dos caracteres morfológicos, previamente distinguidos pela análise de correspondência discriminante, revelou a mesma similaridade entre os taxa, sugerindo a divisão de Hyptis em 2 subgêneros. Procedeu-se também à análise multivariada da composição química dos óleos essenciais de 13 espécies de Hypenia (capítulo 2). Os resultados indicaram a presença de 2 grupos de óleos em relação às seções botânicas das amostras. O primeiro grupo (classe I) incluiu as 3 espécies da seção Densiflorae em adição a H. subrosea Harley e H. aristulata (Epl.) R. Harley, o qual foi caracterizado pelo maior percentual de α-muurolol (5,85 ± 3,08%). Na classe II, com 8 espécies da seção Laxiflorae, os principais constituintes discriminantes foram o (E)-cariofileno (7,09 ± 4,88%), germacreno D (18,1 ± 11,4%) e o biciclogermacreno (6,65 ± 1,19%). Todos os óleos essenciais apresentaram predominantemente sesquiterpenos, tais como espatulenol (4,5-31,6%), óxido de cariofileno (2,2-14,4%) e selin-11-en-4α-ol (0-34,8%). Os agrupamentos foram idênticos quando utilizada a análise multivariada baseada nos esqueletos carbônicos dos constituintes químicos ou utilizando 18 caracteres morfológicos das folhas das espécies.

Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2015-07-28T01:31:42Z No. of bitstreams: 1 Disserta??o Bianca Final.pdf: 956007 bytes, checksum: beb73baaedae8e53837ad84aa8d164cc (MD5) / Made available in DSpace on 2015-07-28T01:31:42Z (GMT). No. of bitstreams: 1 Disserta??o Bianca Final.pdf: 956007 bytes, checksum: beb73baaedae8e53837ad84aa8d164cc (MD5) Previous issue date: 2014-03-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Lamiaceae is a family of flowering plants of great importance due to its aromatic herbs with pharmaceutical properties. Endemic species from the semiarid region of Brazil, as Martianthus leucocaphalus (Mart. Ex Benth.) J. F. B. Pastore, produce essential oils of great phytochemical and economic potential, due to its proven antimicrobial and antifungal activities. This species is not yet commercially cropped, but it is already known that biotic and abiotic factors play an important role on the production of essential oils, making it difficult to to explore these compounds. In this sense, we aimed to evaluate the effect of climate in different months of the year in edaphoclimatic conditions of Feira de Santana, BA, on the growth, production and composition of essential oil from M. leucocaphalus. Experiments were performed on Horto Florestal Experimental Unit from Universidade Estadual de Feira de Santana. We monthly quantified growth, by foliar area parameters, using fresh and dried leaves, branches and inflorescences. In each period we evaluated content and chemical composition of essential oilds under a combination of Gas Chromatography techniques (CG), in order to quantify constituents and Gas Chromatography coupled to Mass Spectrometry (CG/EM) to identify these constituents. Climatic data were obtained over INMET. Obtained data were analyzed under SAS, using Spearman correlation among variables and descriptive statistics. A mean test was performed on SISVAR for comparison. Total dried mass (MST) and essential oil content (TO) were higher on months of great solar intensity and lower on months with abundant precipitation. Constituents varied considerably due to climatic factors, with major concentration of the majoritary compound on more humid months with a low solar irradiance. / A fam?lia Lamiaceae possui not?vel import?ncia devido ?s suas ervas arom?ticas com potencial farmac?utico. Esp?cies end?micas do semi?rido do Brasil, como Martianthus leucocaphalus (Mart. ex Benth.) J.F.B.Pastore produzem ?leo essencial com elevado potencial fitoqu?mico e econ?mico, com atividade antimicrobiana e antif?ngica comprovada. A esp?cie ainda n?o ? cultivada comercialmente, entretanto, sabe-se que a produ??o de ?leos essenciais sofre grande influ?ncia de fatores bi?ticos e abi?ticos, o que dificulta a explora??o desses compostos. Visando avaliar a influ?ncia clim?tica na produ??o do ?leo essencial de M. leucocephalus e, com isso, identificar a melhor ?poca de plantio e colheita da esp?cie para a produ??o de compostos bioativos, o objetivo desse trabalho foi avaliar o efeito do clima, ao longo de diferentes meses do ano, em condi??es edafoclim?ticas de Feira de Santana-BA, sobre o crescimento, produ??o e composi??o do ?leo essencial dessa esp?cie. O experimento foi realizado na Unidade Experimental Horto Florestal da Universidade Estadual de Feira de Santana. Foi quantificado mensalmente o crescimento, atrav?s dos par?metros ?rea foliar, massa fresca e seca de folhas, ramos e infloresc?ncias. Em cada per?odo foi avaliado o teor e composi??o qu?mica dos ?leos essenciais atrav?s da combina??o de t?cnicas de Cromatografia Gasosa (CG), para quantifica??o dos constituintes e Cromatografia Gasosa acoplada a Espectrometria de Massas (CG/EM) para identifica??o dos mesmos. Os dados clim?ticos foram obtidos atrav?s do INMET. Os dados obtidos foram analisados atrav?s da ANAVA, realizando-se a correla??o de Spearman entre as vari?veis e teste de Scott-Knott para compara??o das m?dias. A massa seca total (MST) e o teor de ?leo essencial (TO) foram maiores nos meses com elevada intensidade solar e menor nos meses com muita precipita??o. Os constituintes variaram consideravelmente em fun??o dos fatores clim?ticos, com maior concentra??o do composto majorit?rio nos meses mais ?midos com baixa irradi?ncia solar.

Made available in DSpace on 2015-05-14T12:59:42Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3313855 bytes, checksum: 4725cee09197381bbb8d4bb6107255b2 (MD5) Previous issue date: 2012-11-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The genus includes many species of Cladosporium fungi and opportunistic dematiaceous contaminants being found in diverse environments. Cladosporium carrionii is considered the most important pathogenic species of this genus due to the numerous cases of disease caused by this fungus in the world, being an agent of chromoblastomycosis, feohifomicoses and allergies. The increased resistance to antifungal agents available and their toxicity lead to search for new antifungals, more effective and less toxic. In this context, this study aimed to identify the components of the essential oil of Melissa officinalis L. and investigate their antifungal activity in vitro against strains of Cladosporium carrionii. The chemical composition of the oil was performed by gas chromatography coupled to mass spectrometer (GC-MS). Assays of antifungal activity were performed by microbiological screening by diffusion technique in solid medium; determination of minimum inhibitory concentration (MIC) by microdilution; minimum fungicidal concentration (MFC); measure radial growth at different time intervals; Inhibition of conidial germination and assessment of morphological changes in the presence of the essential oil. The results of GC-MS showed 4 major components, being geranial (52%) the major constituent, followed by the citral (38.90%), trans-&#946;-caryophyllene (1.22%) and germacrene D (0.84%), in descending order of percentage. In antifungal activity assays, the essential oil of M. officinalis inhibited the growth of 100% of the strains of C. carrionii tested, and its MIC and MFC established at 256 mg / mL, and is considered a product with strong antifungal activity. Was able to induce radial mycelial growth inhibition from the concentration of 128 mg / ml (MIC/2), and the concentrations MICX2 and MICX4, there was complete inhibition of the growth. It was also observed significant inhibition of conidial germination in all concentrations compared to control (p <0.05), those potentiated inhibition with increased concentration so that the concentration was MICX2 100% inhibition of germination of conidia counted. Furthermore, morphological changes such as decreased conidiation and structural changes in hyphae were observed in the strains tested, after exposure to oil concentrations MIC/2, MIC and MICX2. Given the foregoing, it is concluded that the essential oil of M. officinalis has a significant antifungal activity against strains of C. carrionii, representing a new possibility in the arsenal of products for treatment of mycoses caused by Cladosporium carrionii. / O gênero Cladosporium abrange muitas espécies de fungos contaminantes e oportunistas dematiáceos, sendo encontrados em diversos ambientes. Cladosporium carrionii é considerada a espécie patogênica mais importante desse gênero, devido aos inúmeros casos de doenças causadas por este fungo em todo o mundo, sendo agente da cromoblastomicose, feo-hifomicoses e quadros alérgicos. O aumento da resistência aos antifúngicos disponíveis e a sua toxicidade levam a busca por novos antifúngicos, mais eficazes e menos tóxicos. Neste contexto, este estudo teve como objetivo identificar os componentes do óleo essencial de Melissa officinalis L. e investigar sua atividade antifúngica, in vitro, sobre cepas de Cladosporium carrionii. A análise da composição química do óleo foi realizada por cromatografia a gás acoplada ao espectrômetro de massas (CG-EM). Os ensaios da atividade antifúngica foram realizados por meio da triagem microbiológica pela técnica de difusão em meio sólido; determinação da concentração inibitória mínima (CIM) pela técnica de microdiluição; concentração fungicida mínima (CFM); medida do crescimento radial em diferentes intervalos de tempo; inibição da germinação de conídios e avaliação das alterações morfológicas na presença do óleo essencial. Os resultados da CG-EM mostraram 4 componentes principais, sendo o geranial (52 %) o constituinte majoritário, seguido do citral (38,90 %), trans &#946;-cariofileno (1,22 %) e germacrene D (0,84 %), em ordem decrescente de percentual. Nos ensaios de atividade antifúngica, o óleo essencial de M. officinalis inibiu o crescimento de 100 % das cepas de C. carrionii ensaiadas, tendo sua CIM e CFM estabelecidas em 256 &#956;g/mL, sendo considerado um produto com forte atividade antifúngica. Foi capaz de induzir inibição do crescimento micelial radial a partir da concentração de 128 &#956;g/mL (CIM/2), e nas concentrações CIMX2 e CIMX4, houve inibição total deste crescimento. Também foi observada inibição significante da germinação de conídios, em todas as concentrações testadas, quando comparados ao controle (p<0,05), sendo estas inibições potencializadas com o aumento da concentração, de modo que na concentração CIMX2 houve inibição de 100% da germinação dos conídios contados. Além disso, alterações morfológicas como diminuição da conidiação e alterações estruturais nas hifas foram observadas nas cepas ensaiadas, após a exposição ao óleo nas concentrações CIM/2, CIM e CIMX2. Diante do exposto, conclui-se que o óleo essencial de M. officinalis apresenta importante atividade antifúngica contra cepas de C. carrionii, representando uma nova possibilidade no arsenal de produtos para terapêutica das micoses causadas por Cladosporium carrionii.

Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-07-05T13:55:07Z No. of bitstreams: 1 EmanoelaPP_TESE.pdf: 1225664 bytes, checksum: 1b71fb8e67155b2279e17cc366f02bd1 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-07-18T15:03:50Z (GMT) No. of bitstreams: 1 EmanoelaPP_TESE.pdf: 1225664 bytes, checksum: 1b71fb8e67155b2279e17cc366f02bd1 (MD5) / Made available in DSpace on 2017-07-18T15:04:02Z (GMT). No. of bitstreams: 1 EmanoelaPP_TESE.pdf: 1225664 bytes, checksum: 1b71fb8e67155b2279e17cc366f02bd1 (MD5) Previous issue date: 2017-02-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Chia (Salvia hispanica L.) is a species that presents considerable economic importance because it is being used as functional foods and by the pharmaceutical industry. The standardization of the germination test of a particular species in the Laboratories of Seeds Analysis, as well as knowing the factors that influence its process, has fundamental importance in the definition of the responses of the species to the intrinsic and extrinsic factors to the seed. In this sense, four experiments with chia seeds were carried out, aiming to determine the appropriate procedures for conducting the germination test and to verify the effects of water and saline stress on the germination and initial development of the seedlings of this species under different temperatures. The experiments were conducted in the Laboratory of Seed Analysis (LAS) of the Federal Rural Semi-Arid University (UFERSA, Mossoró Campus–RN). The statistical design was completely randomized, with four replications. In the first experiment, the influence of two substrates and six levels of wetting on the germination of chia seeds was evaluated. For this, the paper towel substrates (Germitest®) and blotting paper and six volumes of water (1.5; 2.0; 2.5; 3.0; 3.5; 4.0) were multiplied by the dried substrate‟s mass. For the second experiment, the effects of different light regimes and temperatures under the germination of S. hispanica seeds were evaluated using three conditions of light (constant light, dark constant and light / dark alternation with 8 hours of light And 16 h dark) and six temperatures (20; 25; 30; 35; 20-30 and 25-30 °C). In the third experiment, the saline concentrations evaluated were 0.0 (control); 4.5; 9.0; 13.5 and 18.0 dS m-1 and four temperatures: 20, 25, 30 and 20-30 °C. In the fourth experiment, five levels of polyethylene glycol (PEG 6000) (0.0; -0.1; -0.2; -0.3; -0.4 MPa) and four temperatures (20, 25, 30, 20-30 °C). In all the experiments, the germination, vigor and biochemical tests of seedlings were evaluated. The S. hispanica seed germination test should be conducted on the blotting substrate (on paper) with the amount of water between 2.5 and 4.0 times the weight of the dry substrate. The seeds germinated in the blotter paper obtained greater germination growth and accumulation of dry matter of the S. hispanica seedlings in relation to the germinated seedlings in the Germitest® towel paper. The S. hispanica seed germination test can be conducted at constant temperatures of 25 °C and alternating 25-30 °C. The germination of the seeds of S. hispanica is indifferent to the luminosity (neutral photoblastic), however, the greatest growth and accumulation of dry mass occur in the presence of light. Salinity levels greater than 4.5 dS m-1 associated with temperatures of 30 or 20-30 °C affect negatively germination, vigor, growth and biochemical components of seedlings. The temperature of 25 °C provides the best conditions for the development of S. hispanica seedlings. The level of polyethylene glycol of -0.4 MPa for all temperatures studied, and -0.3 MPa at temperatures of 30 °C and 20-30 °C made the germination and vigor of S. hispanica seedlings unfeasible. The S. hispanica seedlings are able to perform osmotic adjustment under water stress conditions up to -0.3MPa when coming from germinated seeds at temperatures up to 25ºC / A chia (Salvia hispanica L.) é uma espécie que apresenta considerável importância econômica, por estar sendo utilizada como alimento funcional e pela indústria farmacêutica. A padronização do teste de germinação de determinada espécie nos laboratórios de Análises de Sementes, assim como conhecer os fatores que influenciam o seu processo, é de fundamental importância na definição das respostas das espécies aos fatores intrínsecos e extrínsecos à semente. Neste sentido, foram realizados quatro experimentos com sementes de chia, objetivando determinar os procedimentos adequados à condução do teste de germinação e verificar os efeitos do estresse hídrico e salino sobre a germinação e desenvolvimento inicial das plântulas desta espécie sob diferentes temperaturas. Os experimentos foram conduzidos no Laboratório de Análise de Sementes (LAS) da Universidade Federal Rural do Semi-Árido (UFERSA, Campus Mossoró - RN). O delineamento estatístico utilizado foi o inteiramente casualizado, com quatro repetições. No primeiro experimento, avaliou-se a influência de dois substratos e seis níveis de umedecimento na germinação de sementes de chia. Para isso, utilizaram-se os substratos papel toalha (Germitest®) e papel mata-borrão e seis volumes de água (1,5; 2,0; 2,5; 3,0; 3,5 e 4,0) vezes a massa do substrato seco. Para o segundo experimento, avaliaram-se os efeitos de diferentes regimes de luz e temperaturas sob a germinação de sementes de S. hispanica, utilizando-se três condições de luminosidade (luz constante, escuro constante e alternância luz/escuro com 8 h de luz e 16 h de escuro) e seis temperaturas (20; 25; 30; 35; 20-30 e 25-30 ºC). No terceiro experimento, as concentrações salinas avaliadas foram as de 0,0 (controle); 4,5; 9,0; 13,5 e 18,0 dS m-1 e quatro temperaturas: 20, 25, 30 e 20-30 °C. No quarto experimento, avaliaram-se cinco níveis de polietilenoglicol (PEG 6000) (0,0; -0,1; -0,2; -0,3 e -0,4 MPa) e quatro temperaturas (20, 25, 30, 20-30 °C). Em todos os experimentos, foram avaliados o teste de germinação, vigor e bioquímicas de plântulas. O teste de germinação de sementes de S. hispanica deve ser conduzido no substrato mata-borrão (sobre papel) com quantidade de água entre 2,5 a 4,0 vezes o peso do substrato seco. As sementes germinadas no papel mata-borrão obtiveram maior germinação crescimento e acúmulo de matéria seca das plântulas de S. hispanica comparadas às plântulas germinadas no papel toalha Germitest®. O teste de germinação de sementes de S. hispanica pode ser conduzido sob temperaturas constante de 25 ºC e alternada de 25-30 ºC. A germinação das sementes de S. hispanica é indiferente à luminosidade (fotoblástica neutra). No entanto, o maior crescimento e acúmulo de massa seca ocorrem na presença de luz. Níveis de salinidade superiores a 4,5 dS m-1, associados às temperaturas de 30 ou 20-30°C, afetam negativamente a germinação, o vigor, o crescimento e os componentes bioquímicos de plântulas. A temperatura de 25 °C proporciona as melhores condições para o desenvolvimento das plântulas de S. hispanica. O nível de polietilenoglicol de -0,4 MPa para todas as temperaturas estudadas e -0,3 MPa nas temperaturas de 30 °C e 20-30 °C inviabilizam a germinação e vigor das plântulas de S. hispanica, que são capazes de realizar ajuste osmótico sob condições de estresse hídrico até os níveis de -0,3MPa, quando provenientes de sementes germinadas em temperaturas de até 25 ºC / 2017-07-05

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The quality of plant-derived drugs must be ensured through tests of authenticity (organoleptic features, macro and microscopic identification), purity and integrity (foreign matter, total ash, acid insoluble ash, loss on drying, microbiological contaminants and heavy metals) and also by qualitative and quantitative determinations of active (s) principle (s) and / or chemical (s) marker (s) characteristic (s) of the species. Many plant species are used in traditional medicine without scientific evidence of its pharmacological properties. The purpose of this work lies in the botanical characterization, physico-chemical and anti-inflammatory activity of Glechon spathulata. Among the macroscopic features of G. spathulata leaves stand out the spatulate form and subrevolute margin, finely crenulate in the middle-upper. Microscopically, the uniseriate epidermis shows straight to slightly sinuous cells, diacytic stomata, and non-glandular and glandular trichomes, dorsiventral mesophyll. Vascular bundles are collateral closed, and no crystals were identified. These features help to the botanical quality control of G. spathulata, and when analyzed in conjunction with its organoleptic properties, allow to ensure the authenticity of the sample. Furthermore, the procedures which measure the extractives content, particle size, volatile oil content, swelling index, bitterness and foaming index, and the chromatographic profile of crude ethanolic extract (CEE), its fractions and essential oil by thin layer chromatography (TLC), allow to verify the authenticity besides to reveal information about purity and integrity of this species. The chromatographic fingerprinting of CEE of G. spathulata by high performance liquid chromatography (HPLC) was carried out in order to determine the nature of the main compounds, and the presumptive identification of the flavonoids rutin and luteolin, also it allowed the verification of the major compound rutin. Besides the authenticity of the extract obtained from aerial parts, the chromatographic fingerprinting of G. spathulata enabled to monitor its quality and to characterize its phenolic compounds. Therefore, the significant anti-inflammatory activity observed by the oral treatment with CEE of G. spathulata in chronic inflammation in rats is probably exerted by polyphenolic compounds. The performed analyses were based on techniques described in literature, official textbooks and Brazilian legislation and were useful in establishing some parameters for the quality control of aerial parts of G. spathulata as raw material for the pharmaceutical and / or food industries. / A qualidade dos derivados de drogas vegetais deve ser assegurada através de testes de autenticidade (caracterização organoléptica, identificação macro e microscópica), pureza e integridade (matéria estranha, cinzas totais, cinzas insolúveis em ácido, perda por dessecação, contaminantes microbiológicos e metais pesados), e ainda por determinações qualitativa e quantitativa do (s) princípio (s) ativo (s) e/ou marcador (es) químico (s) característico (s) da espécie. Muitas espécies vegetais são utilizadas na medicina tradicional sem comprovação científica das suas propriedades farmacológicas. O objetivo deste trabalho reside na caracterização botânica, físico-química e atividade anti-inflamatória de Glechon spathulata. Entre as características macroscópicas das folhas de G. spathulata destacam-se a forma espatulada e a margem subrevoluta, crenada no meio-superior. Microscopicamente, destacam-se a epiderme uniestratificada com células retas a pouco sinuosas, estômatos diacíticos, tricomas tectores e glandulares, mesofilo dorsiventral, feixes vasculares do tipo colateral fechado com bainha parenquimática, que na nervura central é incompleta e constituída por esclerênquima, bem como a ausência de cristais. Estas características contribuem no controle botânico de qualidade de G. spathulata, quando analisadas em conjunto com suas propriedades organolépticas, permitem a autenticidade da amostra. Por outro lado, os teores extrativos, a granulometria, conteúdo de óleos voláteis, índices de intumescência, amargor e espuma, bem como o perfil cromatográfico do extrato bruto (EB), suas frações e do óleo essencial através da cromatografia em camada delgada (CCD), além de permitir a autenticidade revelam informações sobre a pureza e integridade desta espécie. O perfil cromatográfico do EB de G. spathulata estabelecido através da cromatografia líquida de alta eficiência (CLAE) permitiu determinar a natureza dos principais compostos presentes, bem como a identificação presuntiva dos compostos flavonóides rutina e luteolina, evidenciando como composto majoritário a rutina. Além da autenticidade do extrato obtido a partir das partes aéreas, o perfil cromatográfico de G. spathulata permite monitorar a sua qualidade e caracterizar os compostos fenólicos presentes, cujas ações farmacológicas são bem descritas na literatura. Desta forma, a significativa atividade anti-inflamatória verificada pelo tratamento via oral com o EB de G. spathulata na inflamação crônica em ratos, é provavelmente exercida pelos compostos polifenólicos presentes. As análises realizadas basearam-se em técnicas descritas na literatura, em compêndios oficiais e na legislação brasileira, e permitiram estabeler parâmetros úteis para o controle de qualidade das partes aéreas de G. spathulata como matéria-prima para a indústria farmacêutica e/ou de alimentos.

Made available in DSpace on 2015-05-14T13:00:02Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 7083796 bytes, checksum: 2aa6f174655d082d78cc059fc1eef2e0 (MD5) Previous issue date: 2014-03-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Mucormycoses are infections that have high rates of morbidity and mortality. They show high resistance to antifungal agents, and there is a limited therapeutic arsenal currently available, therefore, there is a great need to give priority to testing therapeutic agents for the treatment of mucormycosis. Along this line, the use of essential oils and phytoconstituents has been emphasized as a new therapeutic approach. The objective of this work was to investigate the antifungal activity of the essential oil (EO) of Thymus vulgaris, and its constituents thymol and p-cymene against Rhizopus oryzae and Rhizopus microsporus, through microbiological screening, determination of minimal inhibitory concentration (MICs) and minimal fungicidal concentration (MFCs), effects on mycelial growth and germination of sporangiospores, fungal morphology and interaction with ergosterol. Also was evaluated the preclinical acute toxicity in mice. In microbiological screening the T. vulgaris essential oil showed antifungal potential against resistant strains of R. oryzae.The MIC of EO and thymol varied 128 512 μg/mL, but the MFC of EO and thymol varied 512 1024 μg/mL and 128 1024 μg/mL, respectively. The results also showed that EO and thymol significantly inhibited mycelial development and germination of sporangiospores of both species of Rhizopus. Investigation of the mechanism of antifungal action showed that EO and thymol interact with ergosterol. These data indicate that EO of T. vulgaris and thymol possess strong antifungal activity, which can be related to their interaction with ergosterol, supporting the possible use of these products in the treatment of mucormycosis. In preclinical acute toxicology the doses of 125, 250, 500 and 1000 mg/kg intraperitoneally (i.p.) showed depressive activity on the central nervous system (CNS). In addition to these parameters was observed that the doses of 125 and 250 mg/kg did not change the body and organs weight of the animals, but it was observed change some of the hematological parameters of the mice. The EO showed DL50 of 250 mg/kg for male and 459.6 mg/kg for female; however the thymol showed DL50 of 222.3 mg/kg for male and 1551 mg/kg for female. These data indicate that EO of T. vulgaris and thymol possess strong antifungal activity, which can be related to their interaction with ergosterol. / As mucormicoses são infecções que possuem elevadas taxas de morbidade e mortalidade, limitado arsenal terapêutico, devido a resistência aos antifúngicos. Portanto, existe uma significativa necessidade de priorizar, testar e aplicar melhorias terapêuticas para o tratamento das mucormicoses. É nesse contexto, que os óleos essenciais e fitoconstituintes vem se destacando como uma nova abordagem terapêutica. O objetivo deste trabalho foi investigar a atividade antifúngica in vitro do óleo essencial (OE) de Thymus vulgaris L. e de seus componentes majoritários (timol e p-cimeno) contra Rhizopus oryzae e Rhizopus microsporus, através da triagem microbiológica, da determinação da concentração inibitória mínima (CIM) e fungicida mínima (CFM), avaliação dos efeitos dos fitoconstituintes no crescimento micelial, na germinação dos esporos fúngicos, na morfologia fúngica e interação com ergosterol. Também foi avaliada a toxicidade pré-clínica aguda em camundongos. Na triagem microbiológica o óleo essencial de T. vulgaris apresentou um dos melhores perfis antifúngicos contra cepas resistentes de R. oryzae. A CIM dos produtos variou entre 128-512 μg/mL, já as CFMs do óleo essencial e timol variaram entre 512-1024 μg/mL e 128-1024 μg/mL, respectivamente. Os resultados também mostraram que tanto o OE como o timol inibiram significativamente o desenvolvimento micelial e a germinação de esporos de ambas as espécies de Rhizopus. Em seguida foi mostrado que os produtos testados alteram a morfologia de R. oryzae e R. microsporus. Na investigação do mecanismo de ação antifúngica foi evidenciado que o OE e o timol interagem com o ergosterol, esterol presente na membrana dos fungos. No ensaio toxicológico pré-clínica agudo, as doses de 125, 250, 500 e 1000 mg/kg via intraperitoneal (i.p.) apresentaram atividade depressora do sistema nervoso central (SNC). Adicionalmente a estes parâmetros foi evidenciado que o OE e o timol nas doses de 125 e 250 mg/kg não promoveram alterações significativas na evolução ponderal e peso dos órgãos dos camundongos. Entretanto, ambas as doses das drogas-teste alteram alguns parâmetros hematológicos dos camundongos. Após 72 h de observação o OE apresentou DL50 estimada em 250 mg/kg para camundongos machos e 459,6 mg/kg para as fêmeas. Já o timol apresentou DL50 estimada em 222,3 mg/kg para os machos e 1551 mg/kg para as fêmeas. Estes dados indicam que o óleo essencial de T. vulgaris e timol, apresentam forte atividade antifúngica, que pode estar relacionada com a interação com ergosterol e consequentemente lise de membrana.

O número de pessoas infectadas por protozoários dos gêneros Leishmania e Trypanosoma cruzi, no mundo, permanece preocupante, havendo cerca de 12 milhões de casos de leishmaniose e 7 a 8 milhões referentes à doença de Chagas. Face ao grave quadro destas doenças e às limitações da terapêutica atual, a busca de novos fármacos é urgente. Considerada uma das principais estratégias disponíveis, a pesquisa a partir de espécies vegetais contendo constituintes bioativos é promissora fonte de moléculas potencialmente ativas. Neste contexto, espécies do gênero Lavandula mostraram atividade anti-Leishmania in vitro, tendo motivado a seleção de L. angustifolia (Lamiaceae). Embora tenham sido comprovadas diversas atividades farmacológicas desta espécie, a atividade antiprotozoária frente aos agentes avaliados ainda não havia sido estudada. Além disto, a produção em larga escala e o seu uso bastante difundido no mundo, sobretudo nas indústrias cosméticas, de perfumes e de domissanitários, despertou interesse. O presente trabalho visou à análise da composição do óleo volátil de uma variedade de L. angustifolia aclimatada ao Brasil e à avaliação de sua atividade antiprotozoária in vitro, frente a cepas de Leishmania infantum, L. amazonensis e de Trypanosoma cruzi. A análise do óleo volátil das inflorescências e das folhas, por CG-EM, indicou a predominância de monoterpenos, tendo-se estabelecido análise comparativa entre estes órgãos. De forma geral, o óleo foi inativo frente às espécies de Leishmania, tendo apresentado promissora atividade anti-Trypanosoma. / The number of people infected by Leishmania and Trypanosoma cruzi protozoa around the world has increased and raised concerns that approximately 12 million cases of leishmaniasis and 7 to 8 million cases of Chagas disease have been reported. The alarming statistics involving these diseases and limitations of therapeutics currently in use point to an urgent need for new drugs. Considered one of the main strategies available, research on plants containing bioactive constituents is a promising source of potentially active molecules. Species of Lavandula genus have shown in vitro anti-Leishmania activity and this encouraged the present study on L. angustifolia (Lamiaceae) species. Although several pharmacological activities have been identified, the antiprotozoal activity against these specific agents has not yet been approached. Moreover, the possibility of a large-scale production and its quite comprehensive use worldwide, especially for the manufacturing of cosmetics, perfumes and household sanitizing products have attracted our interest. This study aimed at the analysis of the volatile oil compounds of the L. angustifolia species acclimatized in Brazil as well as the evaluation of the in vitro antiprotozoal activity against the strains of Leishmania (L.) infantum, L. (L.) amazonensis and Trypanosoma cruzi. The CG-EM analysis of the volatile oil from the inflorescences and leaves has shown monoterpenes as majority constituents also allowing a comparative analysis between these organs. Overall, the oil was not active against the Leishmania species but presented promising anti-Trypanosoma activity.

Submitted by Carolina Neves (carolinapon@uefs.br) on 2016-08-10T00:55:37Z No. of bitstreams: 1 Diserta??o_Uiliane.pdf: 2196502 bytes, checksum: 19a832ffbbf5c04770799ccc45403039 (MD5) / Made available in DSpace on 2016-08-10T00:55:37Z (GMT). No. of bitstreams: 1 Diserta??o_Uiliane.pdf: 2196502 bytes, checksum: 19a832ffbbf5c04770799ccc45403039 (MD5) Previous issue date: 2016-03-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The cultures of vine and tomato have an important economic role in Brazil. However phytosanitary issues can compromise the productivity of crops, one issue which stands out are diseases caused by fungi and bacteria that result in immeasurable harm to farmers. Considering the economic, environmental impact and health of the population resulting from the massive use of pesticides it is extremely important that there as studies done to find an alternative control of diseases, such as the use of natural products with antimicrobial activity. Plant extracts and essential oils appear as potential alternatives that can be incorporated into management of phytopathogens, since they can be highly volatile and low persistence, which results in less contamination of the environment. In this context, the Lamiaceae family species is notable for presenting important biological activities, among them high antimicrobial activity. Based on the above, the present study aimed to evaluate the antimicrobial activity of methanol extract and essential oils of species of Eplingiella fruticosa (Salzm. Ex Benth.) Harley & J.F.B.Pastore, Gymneia platanifolia (Mart. ex Benth.) Harley & J.F.B.Pastore and Medusantha martiusii (Benth.) Harley & J.F.B.Pastore against bacteria as Xanthomonas campestris pv. viticola (Naydu) Dye e Ralstonia solanacearum (Smith), causing bacterial canker of grapevine and tomato bacterial wilt, respectively, and fungi causing rots post-harvest vine Aspergilus niger Van Tieghem, Cladosporium herbarum (Pers.:Fr.) Link, Lasiodiplodia theobromae (Pat.) Griffon & Maubl e Rhizopus sp. The plant species were harvested at Feira de Santana, BA. The collected material (leaves) after drying at ambient temperature, were used for the preparation of the extracts (crude methanol), and the extraction of essential oils. The chemical constituents of essential oils were quantified and identified through the techniques of GC/FID and GC/MS. To check the antibacterial activity of extracts and essential oils the diffusion method in agar by the method of discs was used. These were impregnated with essential oils and extracts at different concentrations. The variable studied was the halo diameter of bacterial growth inhibition. For the verification of the minimum inhibitory concentration (MIC) of the extracts and oils microdilution broth method were used. For the evaluation of the antifungal activity diffusion on agar method was used. The methanol extracts and essential oils were incorporated into the PDA culture at different concentrations. The variable analyzed was the diameter of the mycelial growth. The data were submitted to analysis of variance and their means compared by Scott-Knott test at 5% probability of error and the analysis concentrations of regression. According to the obtained results can infer that the methanolic extracts and essential oils of Eplingiella fruticosa, Medusantha martiusii and Gymneia platanifolia species showed antibacterial activity against bacteria X. campestris pv. viticola and R. solanacearum, in two concentrations and MICs ranging from 125 ?L mL-1 to 1000 ?L mL-1, with a strong to moderate antibacterial activity. The methanol extracts and essential oils studied have antifungal activity against phytopathogens such as Aspergilus niger, Cladosporium herbarum, Lasiodiplodia theobromae and Rhizopus sp. / As culturas da videira e do tomateiro possuem importante papel econ?mico no Brasil. Por?m problemas fitossanit?rios podem comprometer a produtividade dessas culturas, entre os quais se destacam doen?as causadas por fungos e bact?rias que causam preju?zos imensur?veis para os agricultores. Considerando o impacto econ?mico, ambiental e ? sa?de da popula??o, decorrentes do uso massivo de defensivos agr?colas, ? de extrema import?ncia estudos visando o controle alternativo de doen?as, como o uso de produtos naturais com atividade antimicrobiana. Os extratos vegetais e os ?leos essenciais surgem como potenciais produtos alternativos que podem ser incorporados ao manejo de fitopat?genos, uma vez que podem ser altamente vol?teis e de baixa persist?ncia, o que resulta em menor contamina??o do ambiente. Neste contexto as esp?cies da fam?lia Lamiaceae destacam-se por apresentar atividades biol?gicas importantes, dentre elas elevada atividade antimicrobiana. Diante do exposto o presente trabalho teve como objetivo avaliar a atividade antimicrobiana do extrato metan?lico e dos ?leos essenciais das esp?cies Eplingiella fruticosa (Salzm. Ex Benth.) Harley & J.F.B.Pastore, Gymneia platanifolia (Mart. ex Benth.) Harley & J.F.B.Pastore e Medusantha martiusii (Benth.) Harley & J.F.B.Pastore contra as bact?rias Xanthomonas campestris pv. viticola (Naydu) Dye e Ralstonia solanacearum (Smith), causadoras do cancro bacteriano da videira e da murcha bacteriana do tomateiro, respectivamente, e dos fungos causadores de podrid?es p?s-colheita da videira Aspergilus niger Van Tieghem, Cladosporium herbarum (Pers.:Fr.) Link, Lasiodiplodia theobromae (Pat.) Griffon & Maubl e Rhizopus sp. A coleta das esp?cies vegetais foi realizada no munic?pio de Feira de Santana - BA. O material coletado (folhas), ap?s a secagem em temperatura ambiente, foi destinado ao preparo dos extratos (metan?lico bruto) e a extra??o de ?leos essenciais. Os constituintes qu?micos dos ?leos essenciais foram quantificados e identificados atrav?s das t?cnicas de CG/DIC e CG/EM. Para a verifica??o da atividade antibacteriana dos extratos e ?leos essenciais foi empregada a metodologia de difus?o em ?gar pelo m?todo de discos. Estes foram impregnados com os ?leos essenciais e extratos em diferentes concentra??es. A vari?vel analisada foi o di?metro do halo de inibi??o do crescimento bacteriano. Para a verifica??o da concentra??o inibit?ria m?nima (CIM) dos extratos e dos ?leos foi utilizado o m?todo de microdilui??o em caldo. Para a avalia??o da atividade antif?ngica foi empregada a metodologia de difus?o em ?gar. Os extratos metan?licos e ?leos essenciais foram incorporados ao meio de cultura BDA em diferentes concentra??es. A var?vel analisada foi o di?metro do crescimento micelial. Os dados obtidos foram submetidos ? an?lise de vari?ncia e as suas m?dias comparadas pelo teste de scott-knott a 5% de probabilidade de erro e para as concentra??es an?lise de regress?o. De acordo com os resultados obtidos pode-se inferir que os ?leos essenciais de E. fruticosa, M. martiusii e G. platanifolia apresentam como compostos majorit?rios o ?xido de cariofileno, espatulenol e E-cariofileno; epi-longipinanol, ?-humuleno e E-cariofileno e espatulenol, ?xido de cariofileno e ?-muurolol, respectivamente. Os extratos metan?licos e ?leos essenciais das esp?cies Eplingiella fruticosa, Medusantha martiusii e Gymneia platanifolia apresentaram atividade antibacteriana contra as bact?rias X. campestris pv. viticola e R. solanacearum, nas diferentes concentra??es utilizadas e as CIMs variaram entre 125 ?L mL-1 a 1000 ?L mL-1. Os extratos metan?licos e ?leos essenciais estudados apresentaram atividade antif?ngica contra os fitopat?genos A. niger, C. herbarum, L. theobromae e Rhizopus sp.

Mentha longifolia subsp. polyadena was collected from seven localities in South Africa and from a single population in Botswana to study the essential oil composition and antimicrobial activity of this ethnomedicinal plant. The essential oils were obtained by hydrodistillation and analysed by gas chromatography coupled to mass spectroscopy (GC/MS) and a cluster analysis was performed on the essential oil dataset. From eight samples (representing eight natural populations), two major chemotypes were identified: (i) a menthofuran rich type (51.4% - 61.6%); and (ii) a cis-piperitone epoxide (14.7% – 35.7%) and piperitenone oxide (14.6% - 65.7%) rich type. The constituent analysis showed quantitative variation with higher amounts of oxygencontaining monoterpenes ranging from 56.5% to 89.6% whilst the sesquiterpene hydrocarbons ranged from 4.4% to 16.7%. The essential oil from the different localities mostly showed moderate to good antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Bacillus cereus, Moraxella catarrhalis, Yersinia enterocolitica and Enterococcus faecalis. The oils were generally inactive against Escherichia coli and Salmonella typhimurium. Candida albicans and Cryptococcus neoformans indicated highest sensitivities for oil samples from Komukwane (3 mg/ml and 0.5 mg/ml respectively) and Prins Albert (0.5 mg/ml and 1.6 mg/ml respectively). The HPLC profiles of the methanol and chloroform (1:1) extracts were more conservative and less variable compared to the essential oils. Two major peaks corresponding to retention times of 22.39 min and 26.47 min were present in all eight samples. Most of the solvent extracts displayed moderate to good antimicrobial activity against Gram-positive pathogens, in particular against S. aureus, S. epidermidis and B. cereus with MIC values ranging from 0.5 mg/ml to 2 mg/ml in most cases. The extracts also demonstrated moderate to good activity against most of the Gram-negative pathogens, in particular against Y. eneterocolitica and M. catarrhalis, with MIC values ranging from 0.5 mg/ml to 2 mg/ml. These results may in part provide scientific evidence for the extensive use of Mentha longifolia in traditional healing.